Abstract
Bone marrow stromal cells (BMSCs) for osteoblast differentiation studies can be obtained by gradient isolation techniques or by directly plating a filtered cell suspension. We compared these two procedures to evaluate whether this step is critical in order to obtain a high number of differentiated colonies. Isolated primary rat BMSCs were cultured in vitro with or without insulin-like growth factor II (IGFII), basic fibroblast growth factor (b-FGF), epidermal growth factor (EGF) or transforming growth factor β1 (TGFβ1), and histochemically and biochemically analysed at different time points. The gradient procedure produced a significantly higher number of colonies capable of osteoblastic differentiation. The growth factors had different effects. In particular, b-FGF and EGF significantly increased the number of Alizarin red S positive colonies, while IGFII and TGFβI exerted inhibitory effects. Nodules obtained on day 21 showed some alkaline phosphatase positive cells and were Von Kossa-positive. These data demonstrate that more differentiated colonies are obtainable from BMSCs isolated by the gradient procedure.
| Original language | English |
|---|---|
| Pages (from-to) | 49-58 |
| Number of pages | 10 |
| Journal | Connective Tissue Research |
| Volume | 42 |
| Issue number | 1 |
| Publication status | Published - 2001 |
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Keywords
- Bone marrow stromal cells
- Differentiation
- Growth factors
- Isolation procedure
- Mineralization
ASJC Scopus subject areas
- Biochemistry
- Immunology
- Nephrology
- Cell Biology
- Orthopedics and Sports Medicine
Cite this
An elevated number of differentiated osteoblast colonies can be obtained from rat bone marrow stromal cells using a gradient isolation procedure. / Lisignoli, G.; Remiddi, G.; Cattini, L.; Cocchini, B.; Zini, N.; Fini, M.; Grassi, F.; Piacentini, A.; Facchini, A.
In: Connective Tissue Research, Vol. 42, No. 1, 2001, p. 49-58.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - An elevated number of differentiated osteoblast colonies can be obtained from rat bone marrow stromal cells using a gradient isolation procedure
AU - Lisignoli, G.
AU - Remiddi, G.
AU - Cattini, L.
AU - Cocchini, B.
AU - Zini, N.
AU - Fini, M.
AU - Grassi, F.
AU - Piacentini, A.
AU - Facchini, A.
PY - 2001
Y1 - 2001
N2 - Bone marrow stromal cells (BMSCs) for osteoblast differentiation studies can be obtained by gradient isolation techniques or by directly plating a filtered cell suspension. We compared these two procedures to evaluate whether this step is critical in order to obtain a high number of differentiated colonies. Isolated primary rat BMSCs were cultured in vitro with or without insulin-like growth factor II (IGFII), basic fibroblast growth factor (b-FGF), epidermal growth factor (EGF) or transforming growth factor β1 (TGFβ1), and histochemically and biochemically analysed at different time points. The gradient procedure produced a significantly higher number of colonies capable of osteoblastic differentiation. The growth factors had different effects. In particular, b-FGF and EGF significantly increased the number of Alizarin red S positive colonies, while IGFII and TGFβI exerted inhibitory effects. Nodules obtained on day 21 showed some alkaline phosphatase positive cells and were Von Kossa-positive. These data demonstrate that more differentiated colonies are obtainable from BMSCs isolated by the gradient procedure.
AB - Bone marrow stromal cells (BMSCs) for osteoblast differentiation studies can be obtained by gradient isolation techniques or by directly plating a filtered cell suspension. We compared these two procedures to evaluate whether this step is critical in order to obtain a high number of differentiated colonies. Isolated primary rat BMSCs were cultured in vitro with or without insulin-like growth factor II (IGFII), basic fibroblast growth factor (b-FGF), epidermal growth factor (EGF) or transforming growth factor β1 (TGFβ1), and histochemically and biochemically analysed at different time points. The gradient procedure produced a significantly higher number of colonies capable of osteoblastic differentiation. The growth factors had different effects. In particular, b-FGF and EGF significantly increased the number of Alizarin red S positive colonies, while IGFII and TGFβI exerted inhibitory effects. Nodules obtained on day 21 showed some alkaline phosphatase positive cells and were Von Kossa-positive. These data demonstrate that more differentiated colonies are obtainable from BMSCs isolated by the gradient procedure.
KW - Bone marrow stromal cells
KW - Differentiation
KW - Growth factors
KW - Isolation procedure
KW - Mineralization
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UR - http://www.scopus.com/inward/citedby.url?scp=0034821037&partnerID=8YFLogxK
M3 - Article
C2 - 11696988
AN - SCOPUS:0034821037
VL - 42
SP - 49
EP - 58
JO - Connective Tissue Research
JF - Connective Tissue Research
SN - 0300-8207
IS - 1
ER -